GSM5730123: CB-derived HSPCs - ASXL1 Trunc - ATAC-seq - Rep1.2; Homo sapiens; ATAC-seq
Nuclei were isolated from 5,000-50,000 cells resuspended in 50 µL of ATAC-seq resuspension buffer (RSB: 10 mM Tris-HCl pH 7.4, 10 mM NaCl, and 3 mM MgCl2 in water) with 0.1% NP40, 0.01% digitonin, and 0.1% Tween-20 and incubated on ice for three minutes. After lysis, 1 mL of ATAC-seq RSB with 0.1% Tween-20 was added and tubes were inverted six times to mix. Isolated nuclei were then spun down. Transposition reactions were incubated at 37°C for 30 minutes in a thermomixer with shaking at 1000 rpm. Reactions were cleaned up using Qiagen MinElute Reaction Cleanup kits and amplified with a target concentration of 20 μl at 4 nM